The work was supported by grants from NIH/NINDS (NS054883), Craig H. Neilsen Foundation (280072), and the Veterans Administration and Canadian Spinal Research Organization. We thank the Rat Resource and Research Center, University of Missouri, Columbia, Missouri, for providing GFP rats.

모든 동물 프로토콜은 기관 동물 관리 및 사용위원회 (IACUC)의 승인을 하였다. 실험실 동물 관리 및 안전에 대한 NIH의 지침을 반드시 따라야한다. 수술 절차와 동물을 적절하게 고통과 불편을 최소화하기위한 처리 하였다. 1 척수 Curgery 및 셀 접목 <ol><li> 모든 수술 이전에 오토 클레이브 수술 도구. 다른 동물에 대한 절차와 악기에서 병원균 및 미생물 오염 물질을 제거 할 수있는 핫 구슬 살균기 (파인 과학 도구)를 사용합니다. 수술시 무균 수술 장갑, 가운 및 커튼을 사용합니다. 각 수술에 대한 무균 수술 기법을 사용한다. </li><li> 암컷 피셔에게 복강 내 주사 (IP)를 통해 투여 케타민 (25 ㎎ / ㎖), 자일 라진 (1.3 ㎎ / ㎖), 및 아세 프로 마진 (0.25 ㎎ / ㎖)의 조합으로 344 래트 (2 ㎖ / kg)을 마취. </li><li> 다시 영역을 면도하고 Betadine 및 에탄올로 반복적으로 피부를 청소합니다.</li><li> # 10 블레이드를 사용하여 피부를 잘라 퉁명스럽게 근육 층을 해부하다. , 척추를 분리 T3 척추를 노출하고, 뾰족한 rongeurs를 사용하여 지느러미 후궁 절제술을 수행하기 위해 메스를 사용합니다. </li><li> 종 방향 및 경막을 절개 척수 덩어리 사이에 약 1mm rostrocaudal 간극을 만드는 홍채 절제술 위 및 microaspiration의 조합을 사용 T4 수준에서 척수를 쪼개다. </li><li> 3-0 Vycril와 완전히 출혈이 중지되며, 봉합 근육까지 2 분, 상처 클립 피부를 닫습니다 -에 대한 일을 기다립니다. </li><li> 락 테이트 링거액 (5 ㎖), 부 프레 노르 핀 (0.035 ㎎ / ㎏), 피하 즉시 수술 후 암피실린 (33 ㎎ / ㎏)을 주입하고 깨어 때까지 따뜻한 인큐베이터에서 쥐를 유지한다. </li><li> 주사 링거와 회 삼일 또는 통증과 고통의 흔적이 사라질 때까지 암피실린 회 십일까지 솔루션 및 부 프레 노르 핀. F까지 다른 동물의 회사에 동물을 반환하지 않습니다ully 복구. </li><li> 수동 비우는 휘어진 방광의 설립까지 두번 약 2 주 동안 하루 방​​광을 비우고, 필요한 경우 다음 생존에 걸쳐 하루에 한번 방광을 비울. </li><li> 2 주 후 전술 한 바와 같이, SCI 래트를 다시 마취 및 척수 병변 사이트를 재 노출. 병변 사이트에서 이식 된 세포를 유지하기 위해 폐쇄 경질을 유지합니다. </li><li> 병변 공동의 진원지 여러 주입 병변의 주동이와 꼬리 인터페이스로, 10 E14 유비쿼터스 GFP 유전자 변형 쥐의 배아에서 수집 피브리노겐에 포함 된 세포 용액 (3.5 × 10 5 μL), ㎕의 트롬빈 5,6 주입 해밀턴 주사기에 연결된 40 μM의 내경으로 뽑아 유리 마이크로 피펫을 사용하는 사이트. </li><li> 근육 층을 봉합하고 상처 클립 피부를 닫습니다. </li><li> 1.8 - 1.6 단계에 설명 된대로 수술 후 주입 및 방광 관리를 수행합니다. </li><li> 한 주 수이물 관류는 척수 7 복강 역행 라벨 교감 신경절 뉴런 0.5 % Fluorogold (FG, 증류수 0.4 ㎖) 주사. </li></ol> 2 송신기 주입 <ol><li> 멸균을 실온에서 2 시간 (최대 10 시간까지) - 적어도 1 transmittersin보기 2 % 글루 타르 알데히드 용액 (증류수 96 ㎖ 중의 50 % glutaraldhyde 4 ml)에 담근다. </li><li> 철저하게 멸균 된 0.9 %의 생리 식염수로 3 회 송신기를 세척하고 사용 (더 이상 일 이상 시간)까지 식염수에 저장합니다. </li><li> 여덟 주 E14 세포 이식 (10 주 포스트 부상) 후, 또는 세포 이식 및 순진한 대조군없이 SCI을 시행 한 쥐를 reanesthetize. </li><li> 복부와 뒷다리를 면도. Betadine로 피부를 청소합니다. 앙와위에서 수술 테이블에 쥐를 놓습니다. </li><li> # 15를 이용하여 블레이드 우측 복부 복부와 허벅 다리의 피부를 절개. </li><li> 의를 통해 잘라ubcutaneous 결합 조직은 작은 가위를 사용하여 대퇴 혈관과 신경의 번들을 노출합니다. </li><li> 곡선 팁 미세 집게를 사용하여 혈관과 신경의 대퇴 동맥을 분리합니다. </li><li> 동맥 아래 세 실크 봉합사를 넣고 각 봉합의 느슨한 매듭을합니다. </li><li> 이후 카테터에 대한 혈관 확장을 유도하는 동맥의 표면에 0.1 ㎖의 리도카인 (2 %)를 적용한다. </li><li> 근위 느슨한 실크 봉합사를 스트레칭에 의해 영구적 인 실크 매듭 및 임시 블록 원심 용기를 고정합니다. </li><li> 20 게이지 바늘을 사용하여 곡선을 천공하고 동맥 카테터 삽입 도구를 사용 측거기 카테터 (8 ㎝ 길이)의 선단을 삽입한다. </li><li> rostrally까지하여 흉부 대동맥에 팁을 배치 4cm에 카테터를 삽입합니다. </li><li> 대퇴 동맥 주위에 세 실크 봉합사를 묶는하여 혈관 내 카테터를 고정합니다. </li><li> 흉곽과 t의 꼬리 가장자리 사이의 측면을 따라 피하 주머니 만들기그는 무딘 가위로 무릎의 범위를 대부분의 두개골 확장. </li><li> 과도한 움직임을 방지하기 위해 송신기 주변 결합 조직에 포켓과 봉합에 송신기 몸체를 삽입한다. </li><li> 제 6 비단실로 피부를 봉합. </li><li> 락 테이트 링거액 (5 ㎖), 부 프레 노르 핀 (0.035 ㎎ / ㎏), 피하 즉시 수술 후 암피실린 (33 ㎎ / ㎏)을 주입하고 깨어 때까지 따뜻한 인큐베이터에서 쥐를 유지한다. </li></ol> 3 기초 평균 동맥압 (MAP) 및 심박수 (HR) 녹화 <ol><li> 일찍이 송신기 주입 다음 일일로, 수신기 패드에 하나의 동물을 넣고에 송신기를 켜십시오. 심장 혈관 매개 변수를 동물을 길들과 안정 15 분 - 약 10 기다립니다. </li><li> 적어도 1 시간 동안 전산 데이터 수집 시스템과 펄스 동맥압로부터 유도되는 기록 쉬고 MAP 및 HR. 데이터마다 5 초를 수집합니다. </li><li> 모니터imals 연속적으로 가시 경련의 발생시 데이터 요소를 제거한다. 각각의 동물의 경우, 평균 데이터 포인트의 평균값을 얻는다. </li></ol> 4 대장 자율 Dysreflexia 팽만에 의한 <ol><li> 송신기 수신기 내부의 음식 펠릿와 함께 제공되는 수건에 NSC-이식 또는 SCI 제어 쥐를 제지합니다. 보통 쥐 절차를 수행하는 동안 협력 머물 수 있습니다. </li><li> 약 2cm의 직장에 라텍스 풍선 팁 카테터를 삽입하고 테이프 팔과 꼬리에 고정. </li><li> 에 송신기의 전원을 켜고 10 기다립니다 - 15 분은 혈압이 preinsertion 기준에 응답 할 수 있습니다. </li><li> 약 30 mmHg로의 압력을 발생하기 위해, 서서히 1 분간 공기 1.4 mL를 10 초 동안 풍선의 팽창에 의해 결장 팽창을 유도한다. </li><li> 기록 MAP 및 전 HR 1 분 동안 1 분, 대장 팽창 후 1 분; 샘플 데이터를 3 분의 절차를 수행하는 동안 3 초마다. </li><li>이 시험 사이에 적어도 15 분 복구 간격으로 동물 당 3 시험 - 2를 수행합니다. </li><li> 만약 더 이상의 평가 위에서 설명한 두 번 투여 마취 조합으로 과다 복용 동물 (IP). 4 % 파라 포름 알데히드이어서 식염수 동물들을 관류. </li><li> 각각의 동물의 경우, 이전에 각각 결장 팽만 동안 값을 평균; 기준 및 각 시험에 대한 팽만에 의한 MAP와 HR의 변화 사이의 차이를 계산; 2 평균 이상 - 3 시험은 평균값을 얻었다. </li></ol> 5 송신기 청소 <ol><li> 마취 후와 관류하기 전에 동물의 몸에서 송신기를 제거합니다. 청소 때까지 증류수로 가득 비커에 즉시 담근다; 텔레 장치의 건조를 방지. </li><li> 실온에서 24 시간 동안 1 % Terg-A-Zyme 효소 세척 용액 (10g / L의 물)로 텔레을 전송합니다. </li><li> 무딘 30 게이지 바늘로 송신기 카테터의 끝을 Regelregel 주사기에 연결. </li><li> 조심스럽게 접힌 부드러운 티슈를 사용하여 송신기를 건조하고 원래 플라스틱 트레이에 보관합니다. </li></ol>

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저자가 공개하는 게 없다.

전통적으로, 유체가 채워진 뉼러 동맥에 삽입하고, 각 동물 11 스냅 단자로서 심혈관 매개 변수를 기록하기 위해 압력 변환기에 연결된다. 연속적으로 장시간 심혈관 성능을 모니터링하는, 무선 원격 측정 시스템은 여러 실험실에서 사용된다. 이 세련 도구는 자유롭게 동물을 이동, 의식의 혈압을 기록 할 수 있습니다. 유체 가득 카테터에 비해 원격 측정은 정확하게 SCI 동물에서 심장 혈?...

심장 혈관 장애는 높은 수준에서 척수 손상 (SCI) 이후에 발생합니다. 이 무질서 혈압과 심박 휴지, 기립 성 저혈압, 운동 유발 성 저혈압 및 부상 레벨 아래 1,2 감각 자극에 응답하여 고혈압과 압력 반사 매개 서맥 에피소드 특징 자율 dysreflexia에 명시된다. 이러한 현상은 척수 손상 환자의 일상 생활을 방해. 따라서, SCI 및 실험 동물의 치료와 심장 혈관 변화의 조사를 위해 효과적인 도구를 확립하는 것이 중요하다. 동물에서 심혈관 기능을 조사하기 위해, 여러 가지 기술들은 혈압과 심박을 모니터링하는데 사용되어왔다. 비침 꼬리 커프스 말초 혈압 3을 측정하기 위해 이용 될 수있는 반면 중앙 심혈관 파라미터는, 캐 뉼러 삽입 및 원격 측정에 의해 기록 될 수있다. 다른 방법에 비해 텔레metry가 자유롭게 움직이는 동물 및 심혈관 기능 (4)의 장기 모니터링 연속 기록 가능 주요 장점을 갖는다. SCI의 동물 모델에서 실험적 자극 후 말초 혈압의 변화를 검출 할 수있을만큼 크지 않을 수있다. 따라서, 적합한 심장 모니터링 기술은 SCI와 동물 선정되어야한다. 본 연구에서, 무선 원격 측정 시스템은 완전한 척수 절개 후 성인 쥐 심장 기능을 모니터링하기 위해 도입되었다. 쥐 병변 사이트에서 동계 쥐 배아 일 14 (E14) 뇌간 유래 신경 줄기 세포 (BS-NSCs의)의 이식을 받았다. 부상없이 이식하고 순진한, 손상되지 않은 쥐와 쥐 컨트롤 역임했다. 원격 측정 절차는 송신기 및 멸균 주입 (도 1), 기저 심혈 관계 파라미터의 기록, 결장 팽만 유도 응답 및 전송기 청소 등을 포함저장.

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a radio-telemetric system to monitor cardiovascular function in rats with spinal cord transection and embryonic neural stem cell grafts

높은 흉추 또는 경추 척수 손상 (SCI)는 심장 혈관 기능 장애로 이어질 수 있습니다. 심혈관 매개 변수를 모니터하기 위해 또는 녹색 형광 단백질을 발현하는 배아 뇌간 유래의 신경 줄기 세포의 그라프없이 T4 척수 절개를받은 쥐의 대퇴 동맥에 무선 송신기에 연결된 카테터를 주입. 이러한 캐뉼라 삽입 또는 꼬리 - 커프와 같은 다른 방법에 비해, 원격 측정은 연속적으로 자유롭게 움직이는 동물에서 혈압 및 심박수를 모니터링하는 것이 유리하다. 또한 장기적으로 다수의 데이터 획득이 가능하다. 척수 손상 쥐에서 대장 팽창 님의 억제되지 않은 상태와 자율 dysreflexia에서 기저 심혈관 데이터가 성공적으로 기록되었다. 송신기는 척수 절개 전에 주입되는 경우 또한, 전 및 후의 SCI 심혈관 파라미터가 동일한 쥐에 비교 될 수있다. 설명 telemet의 한 가지 제한RY 과정은 대퇴 동맥에 주입이 동측 뒷다리에 혈액 공급에 영향을 미칠 수 있다는 것이다.

상기 원격 측정 기술을 이용하여, 우리는 성공적 척수 손상 동물에서 심혈관 매개 변수를 기록했다. HR 이전 보고서 9과 일치 순진한 동물에 비해 증가하는 반면 단독으로 SCI와 동물에서 MAP은 크게 감소 하였다. BS-NSC 이식,지도 및 HR과 동물에서 순진한 동물에서 측정 된 수준 (그림 2)에 접근했다. 유해 자극 MAP 상승 및 3,8 HR의 감소를 생성하면 결장 팽만 동안, 래트는 dy...

Watch this Scientific Journal Video about A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts at JoVE.com

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

우리는 병변 부위에 이식 팔주 배아 뇌간 신경 줄기 세포 후 T4 척수 가로로 쥐 심혈관 매개 변수를 기록하기 위해 무선 원격 측정 시스템을 사용하기위한 프로토콜을 제시한다. 텔레 메 트리 정확하게 의식이 자유롭게 이동 척수 손상 쥐의 심장 기능을 평가하는 첨단 기술이다.

무선 원격 측정 시스템은 척수 절개 및 배아 신경 세포 이식 줄기와 흰쥐의 심장 혈관 기능을 모니터링 할 수

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

a-radio-telemetric-system-to-monitor-cardiovascular-function-rats

Research

JoVE Journal

Medicine

10.4K 조회수.  Drexel University College of Medicine.  우리는 병변 부위에 이식 팔주 배아 뇌간 신경 줄기 세포 후 T4 척수 가로로 쥐 심혈관 매개 변수를 기록하기 위해 무선 원격 측정 시스템을 사용하기위한 프로토콜을 제시한다. 텔레 메 트리 정확하게 의식이 자유롭게 이동 척수 손상 쥐의 심장 기능을 평가하는 첨단 기술이다. 

비디오: A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

Assessing Endothelial Vasodilator Function with the Endo-PAT 2000

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of the panoply of paracrine factors that it secretes, the endothelium regulates the contractile and proliferative state of the underlying vascular smooth muscle, as well as the interaction of the vessel wall with circulating blood elements. Because of its central role in mediating vessel tone and growth, its position as gateway to circulating immune cells, and its local regulation of hemostasis and coagulation, the the properly functioning endothelium is the key to cardiovascular health. Conversely, the earliest disorder in most vascular diseases is endothelial dysfunction.
In the arterial circulation, the healthy endothelium generally exerts a vasodilator influence on the vascular smooth muscle. There are a number of methods to assess endothelial vasodilator function. The Endo-PAT 2000 is a new device that is used to assess endothelial vasodilator function in a rapid and non-invasive fashion. Unlike the commonly used technique of duplex ultra-sonography to assess flow-mediated vasodilation, it is totally non-operator-dependent, and the equipment is an order of magnitude less expensive. The device records endothelium-mediated changes in the digital pulse waveform known as the PAT ( peripheral Arterial Tone) signal, measured with a pair of novel modified plethysmographic probes situated on the finger index of each hand. Endothelium-mediated changes in the PAT signal are elicited by creating a downstream hyperemic response. Hyperemia is induced by occluding blood flow through the brachial artery for 5 minutes using an inflatable cuff on one hand. The response to reactive hyperemia is calculated automatically by the system. A PAT ratio is created using the post and pre occlusion values. These values are normalized to measurements from the contra-lateral arm, which serves as control for non-endothelial dependent systemic effects. Most notably, this normalization controls for fluctuations in sympathetic nerve outflow that may induce changes in peripheral arterial tone that are superimposed on the hyperemic response.
In this video we demonstrate how to use the Endo-PAT 2000 to perform a clinically relevant assessment of endothelial vasodilator function.

A noninvasive procedure to assess endothelial function is demonstrated using the Endo-PAT 2000.

엔도 - PAT 2000 내피 Vasodilator 기능을 평가

The endothelium is a delicate monolayer of cells that lines all blood vessels, and which comprises the systemic and lymphatic capillaries. By virtue of ...

연구

의학

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

쥐의 척수를 다친 후에 급성 및 만성 촉각 감각 테스트

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

쥐에 심각한 척수 손상의 타박상 모델

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

실험 경추 척수 손상의 신경 전구체 세포 및 자기 조립 펩티드의 시너지 사용

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

척수 손상의 실험 Contusive 모델에서 신경 줄기 세포 이식

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

쥐의 척수 손상의 모델로 초점 국소 빈혈을 Photothrombosis는 유도

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

신생아 마우스 척수 압축 부상 모델

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

정맥 삽입 위치 모델 : 바이 패스 이식 개통을 연구하기위한 적절한 모델

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Studying the Hypothalamic Insulin Signal to Peripheral Glucose Intolerance with a Continuous Drug Infusion System into the Mouse Brain

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues contributes to type 2 diabetes mellitus (T2DM) development and appetite regulation in the hypothalamus. Chemokine CCL5 and C-C chemokine receptor type 5 (CCR5) levels have been suggested to mediate arteriosclerosis and glucose intolerance in type 2 diabetes mellitus (T2DM). In addition, CCL5 plays a neuroendocrine role in the hypothalamus by regulating food intake and body temperature, thus, prompting us to investigate its function in hypothalamic insulin signaling and the regulation of peripheral glucose metabolism.
The micro-osmotic pump brain infusion system is a quick and precise way to manipulate CCL5 function and study its effect in the brain. It also provides a convenient alternative approach to generating a transgenic knockout animal. In this system, CCL5 signaling was blocked by intracerebroventricular (ICV) infusion of its antagonist, MetCCL5, using a micro-osmotic pump. The peripheral glucose metabolism and insulin responsiveness was detected by the Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT). Insulin signaling activity was then analyzed by protein blot from tissue samples derived from the animals.
After 7-14 days of MetCCL5 infusion, the glucose metabolism and insulin responsiveness was impaired in mice, as seen in the results of the OGTT and ITT. The IRS-1 serine302 phosphorylation was increased and the Akt activity was reduced in mice hypothalamic neurons following CCL5 inhibition. Altogether, our data suggest that blocking CCL5 in the mouse brain increases the phosphorylation of IRS-1 S302 and interrupts hypothalamic insulin signaling, leading to a decrease in insulin function in peripheral tissues as well as the impairment of glucose metabolism.

This protocol studies the role of chemokine (C-C motif) ligand 5 (CCL5) in the hypothalamus by delivering an antagonist, MetCCL5, into the mouse brain using a micro-osmotic pump brain infusion system. This transient inhibition of CCL5 activity interrupted hypothalamic insulin signaling, leading to glucose intolerance and peripheral systemic insulin sensitivity.

Insulin regulates systematic metabolism in the hypothalamus and the peripheral insulin response. An inflammatory reaction in peripheral adipose tissues ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

풍선-기반 부상 마우스 복 부 대동맥에서 Myointimal 증식을 유도 하

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...